In recent years, analysis of various components, genetic diagnosis and the like are performed by measurement of fluorescence, absorbance or reflectance. For example, in component analysis using fluorescence measurement, a sample mixed with a coloring matter (fluorescent coloring matter) is irradiated with light, and an intensity of fluorescence excited by the light is measured to detect a material labeled with the coloring matter (fluorescent coloring matter).
In component analysis using absorbance measurement, a sample mixed with a coloring matter is irradiated with light having a wavelength corresponding to the coloring matter, and an intensity of transmitted light is measured to calculate absorbance, thereby detecting a material labeled with the coloring matter, as disclosed in, for example, JP No. 9-21749A. In component analysis using reflectance measurement, an intensity of scattered light, instead of transmitted light, is measured to calculate reflectance, thereby detecting a material labeled with the coloring matter.
In the case of detection of a plurality of materials using the above-described component analysis, a sample is mixed with a plurality of different coloring matters that vary depending on the materials to be detected, and the sample is irradiated with light corresponding to each coloring matter separately.
In the case of fluorescence measurement, component analysis is performed by irradiating a sample mixed with a plurality of coloring matters (fluorescent coloring matters) that have different excitation wavelengths and fluorescence wavelengths, with light having the excitation wavelength of each coloring matter separately, and measuring a fluorescence intensity of the coloring matter, as disclosed in, for example, JP 2000-503774A.
In the case of absorbance measurement, component analysis is performed by irradiating a sample mixed with a plurality of coloring matters having different absorption wavelengths, with light having the absorption wavelength of each coloring matter separately, to measure an intensity of transmitted light for each coloring matter.
However, in general, the excitation wavelength, absorption wavelength and reflection wavelength of a coloring matter have a certain width. Therefore, in fluorescence measurement, if the coloring matters (fluorescent coloring matters) used have close excitation peak wavelengths, when a certain coloring matter is excited by light with its excitation wavelength, other coloring matter(s) also may be excited by the light. In this case, the resultant fluorescence intensity is a value obtained by combining the fluorescence intensity of each excited coloring matter, thereby making it difficult to perform accurate component analysis, genetic diagnosis or the like.
The same is true of absorbance measurement and reflectance measurement. Specifically, the resultant intensity of transmitted light or scattered light is a value obtained by combining the intensity of transmitted light or scattered light for each coloring matter, thereby making it difficult to perform accurate component analysis, genetic diagnosis or the like.
An object of the present invention is to provide a measuring instrument and a fluorometric method capable of separating and measuring an actual intensity of each coloring matter from a combined value of the intensity of transmitted light or radiated light.